The goal of this project is to identify and validate interstitial cystitis/painful bladder syndrome (IC/PBS)- associated urinary metabolites. IC/PBS is a debilitating condition that presents with a constellation of symptoms including bladder pain, urinary urgency, frequency, nocturia, and small voided volumes in the absence of other identifiable etiologies. The diagnosis of IC/PBS remains dependent on subjective parameters, leading to extreme difficulties in accurately phenotyping patients. Our central hypothesis is that IC/PBS- associated metabolites in the urine of IC/PBS patients can segregate patients from control subjects, and that their levels are correlated with clinical symptoms. This hypothesis is based on our proton nuclear magnetic resonance (NMR) spectroscopy findings identifying urine metabolites that appear to stratify IC/PBS patients from healthy controls. We propose to substantially build on these results and determine whether candidate urine metabolites are diagnostic indicators of IC/PBS. To test this hypothesis we will (Aim 1) Identify IC/PBS-associated metabolites in urine using two independent platforms, proton NMR spectroscopy and quadrupole time-of-flight (Q-TOF) mass spectrometry. We will determine whether candidate metabolites segregate IC/PBS patients from control subjects, and are associated with clinical severity, including chronic bladder pain. In order to achieve a deeper understanding of the underlying biological nature of this condition, we will identify pathologic networks associated with IC/PBS using integrative bioinformatics. We will also (Aim 2) create a Cedars-Sinai Medical Center IC/PBS urine biorepository and will conduct preliminary validation studies of IC/PBS metabolites. Our long-term goals include the identification of diagnostic urinary biomarkers that can serve as non-invasive and accurate means of diagnosing IC/PBS and stratifying these patients from patients with other bladder or pelvic floor conditions. We also seek in the long term to validate molecular targets for therapeutic strategies. This study has a significant potential clinical impact because results may lead to clinical methods to increase diagnostic accuracy and an improved understanding of the molecular basis of IC/PBS and its relationship to urologic conditions with overlapping symptoms.